GenMute™ siRNA Transfection Reagent

Description:
GenMute™ Reagent is a novel biodegradable non-liposomal siRNA/miRNA & DNA delivery tool. With our proprietary pH Dependent Conformational Change (PDCC) technology, the biodegradable transfection reagent was formulated by addition of pre-screened hydrophobic groups to its side chain, making GenMute™ Reagent a versatile and most powerful gene delivery tool in the market. GenMute™ Reagent has been validated to effectively and reproducibly transfect single siRNA/miRNA or co-transfect DNA/siRNA or DNA/miRNA to variety of mammalian cells.

Size & content:
– GenMute™ Reagent, 1.0 mL, sufficient for ~2,000 reactions based on transfecting 5.0 pmol siRNA or miRNA mimics in a 24-well plate.
– GenMute™ Transfection Buffer (5x ), formulated for maximal transfection efficiency, 8.0 mL to make 40 mL working solution.

Applications:
– Single siRNA transfection or DNA/siRNA co-transfection to a variety of mammalian cells.
– Single miRNA, miRNA mimics or inhibitors transfection or DNA/miRNA or DNA/miRNA mimics co-transfection to a variety of mammalian cells.

Storage:
Store at 4 °C for GenMute™ Reagent and RT for GenMute™ Transfection Buffer (5x ). If stored properly, the product is stable for 12 months or longer.

Advantages:
– Excellent silencing at low concentrations with only 1.0 nM siRNA / miRNA mimics
– Minimizing non-specific and off-target effects in the cell
– One tube reaction
– Best for broad mammalian cells
– Very low cytotoxicity
– Very affordable

Comparisons of Silencing Efficacy of GenMute™ siRNA & DNA Transfection Reagent with Brand Name Products

Knockdown efficacy comparison of GenMute™ Transfection Reagent (upper panel) vs. Dharmafect™ 4 siRNA Transfection Reagent (middle panel) and Lipofectamine™ 2000 (lower panel) on HEK293 cells. siRNA targeting renilla luciferase at different final concentrations ranging from 0.5 to 20 nM was co-transfected with the renilla luciferase gene (0.5 µg of pRL-CMV DNA per well) by the above three transfection reagents per manufacturer’s protocols into HEK293 cells growing on a 24-well plate. Renilla luciferase activity was determined 24h after post co-transfection with the renilla luciferase determination system (Promega). The luminescence was measured from 5.0 µl of lysate during 10s integration with a luminometer (Beckman Coulter LD 400). Luciferase activity was expressed as light units integrated over 10s (RLU) and normalized per mg of cell protein by using the BCA assay. The error bars represent the standard deviation derived from triplicate experiments. Luciferase-silencing efficiency was calculated relative to untreated cells. While GenMute™ and Dharmafect™ 4 reagents delivered significant gene silencing from 1.0 nM of renilla luciferase siRNA, lipofectamine™ 2000 gave good knockdown only after 30 nM (data not shown).

Excellent silencing of endogenously expressed KIF11 (also known as Eg5) in HEK293 cells with 0.5 µl of GenMute™ reagent and 5.0 pmol Eg5 siRNA per well of 24-well plate. KIF11 (also known as Eg5) encodes a motor protein that belongs to the kinesin-like protein family involved in chromosome positioning and bipolar spindle formation during cell mitosis. A reduction in KIF11 levels causes mitotic arrest. GenMute™ reagent effectively delivers Eg5 siRNA (final 10 nM, right panel) to HEK293 cells at only 0.5 µl per well of 24-well plate vs. negative control (final 10 nM. left panel). Compared with the negative control (left panel), the phenotype of “rounded-up” 293 cells were visualized 24 hours post transfection (right panel) with a Nikon microscope.

GenMute™ Transfection Reagent knocked down endogenous lamin A/C gene expression in HeLa cells. A siRNA targeting lamin A/C gene (right panel) and a sham siRNA (left panel) were introduced into Hela cells (final 1.0 nM) by GenMute™ Transfection Reagent. Lamin A/C gene silencing was monitored 24h post-transfection by immunofluorescence. Lamin A/C was probed with a mouse monoclonal lamin A/C specific antibody, followed by the addition of FITC-conjugated anti-mouse antibodies. Quantitative analysis showed that lamin siRNA at 1.0 nM delivered by GenMute™ Transfection Reagent knocked down 94% endogenously expressed lamin A/C in Hela cells.

Data Sheet & Protocol
– A Protocol for siRNA Transfection and DNA/siRNA Co-Transfection
– A Short Protocol for siRNA Transfection for Advanced Users 
– A Short Protocol for siRNA/DNA Co-transfection for Advanced Users 
– A Protocol for DNA Transfection to Mammalian Cells